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|Title:||High affinity nucleic acid ligands containing modified nucleotides|
|Abstract:||Nucleic acid ligands containing modified nucleotides are described as are methods for producing such oligonucleotides. Such ligands enrich the chemical diversity of the candidate mixture for the SELEX process. Specific examples are provided of nucleic acids containing nucleotides modified at the 2'- and 5-position. Specific 2-OH and 2'-NH.sub.2 modified RNA ligands to thrombin are described.|
|Inventor(s):||Pieken; Wolfgang (Longmont, CO), Tasset; Diane (Boulder, CO), Janjic; Nebojsa (Lafayette, CO), Gold; Larry (Boulder, CO), Kirschenheuter; Gary P (Arvada, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Sep 27, 1999|
|Claims:||1. A method of preparing 5-iodo,2'-amino deoxyuridine from uridine comprising: |
a) dehydrating uridine to 2,2'-anhydrouridine;
b) converting 2,2'-anhydrouridine to an unprotected 2'-azido,2'-deoxyuridine;
c) iodinating the unprotected 2'-azido,2'-deoxyuridine to 5-iodo, 2'-azido,2'-deoxyuridine; and
d) reducing the unprotected 5-iodo,2'-azido,2'-deoxyuridine to 5-iodo,2'-amino,2'-deoxyuridine.
2. A method of preparing 5-iodo,2'-amino,2'-deoxy uridine 5'-triphosphate from 2'-amino,2'-deoxyuridine, comprising:
a) trifluoroacetylating 2'-amino,2'-deoxyuridine to form 2'-trifluoroacetylamino,2'-deoxyuridine;
b) phosphorylating 2'-trifluoroacetylamino,2'-deoxyuridine to 2'-amino,2'-deoxyuridine 5'-triphosphate; and
c) iodinating 2'-trifluoroacetylamino,2'-deoxyuridine by mercuration to form 5'-iodo,2'-amino,2'-deoxyuridine 5'-triphosphate.
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